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To obtain more information of Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) transmission via faeces in/between farms, 360 swine faecal samples were randomly collected from different farms in China from 2017 to 2019. Sixty‐two ORF5 genes were amplified by PCR from 120 positive samples identified by real‐time RT‐PCR and further characterized by sequencing. Phylogenetic analysis based on the ORF5 gene revealed that these strains can be divided into four lineages: lineage 1 (NADC30‐like), lineage 3 (QYYZ‐like), lineage 5.1 (VR2332‐like) and lineage 8.7 (JXA1‐like), with 62.9% (39/62) NADC30‐like virus, 21% (13/62) QYYZ‐like virus, 1.6% (1/62) VR2332‐like virus and 14.5% (9/62) for JAX1‐like virus. In particular, 14 PRRSVs including lineage 1, 5.1 and 8.7 can be isolated from 120 positive faecal samples, which further suggests that faecal transmission may be an important factor in the spread of PRRSV in farms. Full‐length genome sequencing analysis showed that 14 isolates share 83.1%–97.7% homology with each other and 82.3%–96.1% identity with NADC30, 83.2%–99.7% with VR2332, 79.6%–87.2% with QYYZ and 82.6%–98.9% with JXA1 and CH‐1a, and only 60.1%–60.7% with LV. Recombination events were observed in the six out of 14 strains. Collectively, the data of this study are useful for understanding the spread of PRRSV via faeces. Additionally, the virus was isolated from positive faecal samples, suggesting that faecal transmission may be an important factor in the spread of PRRSV in farms. [ABSTRACT FROM AUTHOR]

Endometritis is a postnatal reproductive disorder disease, which leads to great economic losses for the modern dairy industry. Emerging evidence indicates that microRNAs (miRNAs) play a pivotal role in a variety of diseases and have been identified as critical regulators of the innate immune response. Recent miRNome profile analysis revealed an altered expression level of miR‐148a in cows with endometritis. Therefore, the present study aims to investigate the regulatory role of miR‐148a in the innate immune response involved in endometritis and estimate its potential therapeutic value. Here, we found that miR‐148a expression in lipopolysaccharide (LPS)‐stimulated endometrial epithelial cells was significantly decreased. Our results also showed that overexpression of miR‐148a using agomiR markedly reduced the production of pro‐inflammatory cytokines, such as IL‐1β and TNF‐α. Moreover, overexpression of miR‐148a also suppressed NF‐κB p65 activation by targeting the TLR4‐mediated pathway. Subsequently, we further verified that miR‐148a repressed TLR4 expression by binding to the 3′‐UTR of TLR4 mRNA. Additionally, an experimental mouse endometritis model was employed to evaluate the therapeutic value of miR‐148a. In vivo studies suggested that up‐regulation of miR‐148a alleviated the inflammatory conditions in the uterus as evidenced by H&E staining, qPCR and Western blot assays, while inhibition of miR‐148a had inverse effects. Collectively, pharmacologic stabilization of miR‐148a represents a novel therapy for endometritis and other inflammation‐related diseases. [ABSTRACT FROM AUTHOR]

Recombinant PRRSV △2ORF5 gene was constructed using DNA shuffling from four genetically different strains of PRRSV to study its heterologous cross-neutralizing ability. The △2ORF5 mutant gene was cloned into the vector pET-32a and transferred into E. coli BL21. SDS-PAGE confirmed that the molecular weight of the recombinant △2ORF5 was about 42 kDa, consistent with the predicted result. Then the purified recombinant protein was injected into BALB/c mouse to obtain polyclonal antibody. Western blotting analysis with mouse-anti-△2ORF5 polyclonal serum indicated that the parental virus recombinant GP5 protein reacted with the specific antibodies. Four parental viruses could be inhibited by the anti-△2ORF5 polyclonal antibody and the inhibition rates were higher than 53%. This work has laid a foundation for further development vaccine for PRRSV.